Rapid Assessment of Tinnitus Complaints with a Modified Version of the Tinnitus and Hearing Survey.

The objective of this project was to establish cutoff scores on the tinnitus subscale of the Tinnitus and Hearing Survey (THS) using a large sample of United States service members (SM) with the end goal of guiding clinical referrals for tinnitus evaluation. A total of 4,589 SM undergoing annual audiometric surveillance were prospectively recruited to complete the THS tinnitus subscale (THS-T). A subset of 1,304 participants also completed the Tinnitus Functional Index (TFI). The original 5-point response scale of the THS (THS-T16) was modified to an 11-point scale (THS-T40) for some participants, to align with the response scale of the TFI. Age, sex, hearing loss, and self-reported tinnitus bother were also recorded. The THS-T was relatively insensitive to hearing, but self-reported bothersome tinnitus was significantly associated with the THS-T40 score. Receiver operating characteristic analysis was used to determine cutoff scores on the THS-T that aligned with recommended cutoff values for clinical intervention on the TFI. A cutoff of 9 on the THS-T40 aligns with a TFI cutoff of 25, indicating a patient may need intervention for tinnitus. A cutoff of 15 aligns with a TFI cutoff of 50, indicating that more aggressive intervention for tinnitus is warranted. The THS-T is a viable tool to identify patients with tinnitus complaints warranting clinical evaluation for use by hearing conservation programs and primary care clinics. The THS-T40 cutoff scores of 9 and 15 provide clinical reference points to guide referrals to audiology.

Examining the Use and Benefits of Low-/Mild-Gain Hearing Aids in Service Members with Normal Hearing Thresholds and Self-Reported Hearing Difficulties.

Low- (or mild-) gain hearing aids (LGHAs) are increasingly considered for individuals with normal peripheral hearing but significant self-reported hearing difficulties (SHDs). This study assesses the benefits of LGHAs as a management option for individuals with normal hearing thresholds (NHTs) and SHDs, comparing LGHA use and benefit to individuals with non-significant hearing difficulties (NHDs) and those with peripheral hearing loss. Questionnaires addressing hearing aid usage, benefit, hearing difficulties, and tinnitus were administered to 186 individuals who self-identified as hearing aid users in a sample of 6652 service members who were receiving their annual hearing tests. Participants were divided into SHD and NHD groups based on the normative cutoff of the Tinnitus and Hearing Survey-Hearing Subscale (THS-H), and into hearing impairment (HI) and NHT based on their audiometric air-conduction thresholds. Individuals with SHDs and NHTs reported higher LGHA usage and benefit than individuals with NHDs and NHTs. Comparable use and benefit were noted between groups with SHDs regardless of peripheral hearing loss status. The findings support LGHAs as a suitable management option for individuals with NHTs and SHDs, as indicated by hearing aid use and benefit. Quantifying the level of perceived auditory processing deficits (i.e., SHDs), notably with the THS-H, enhances sensitivity in identifying those who may benefit the most from this treatment option.

The Effects of Signal to Noise Ratio, T60 , Wide-Dynamic Range Compression Speed, and Digital Noise Reduction in a Virtual Restaurant Setting.

OBJECTIVES: Hearing aid processing in realistic listening environments is difficult to study effectively. Often the environment is unpredictable or unknown, such as in wearable aid trials with subjective report by the wearer. Some laboratory experiments create listening environments to exert tight experimental control, but those environments are often limited by physical space, a small number of sound sources, or room absorptive properties. Simulation techniques bridge this gap by providing greater experimental control over listening environments, effectively bringing aspects of the real-world into the laboratory. This project used simulation to study the effects of wide-dynamic range compression (WDRC) and digital noise reduction (DNR) on speech intelligibility in a reverberant environment with six spatialized competing talkers. The primary objective of this study was to determine the efficacy of WDRC and DNR in a complex listening environment using virtual auditory space techniques. DESIGN: Participants of greatest interest were listeners with hearing impairment. A group of listeners with clinically normal hearing was included to assess the effects of the simulation absent the complex effects of hearing loss. Virtual auditory space techniques were used to simulate a small restaurant listening environment with two different reverberation times (0.8 and 1.8 sec) in a range of signal to noise ratios (SNRs) (-8.5 to 11.5 dB SNR). Six spatialized competing talkers were included to further enhance realism. A hearing aid simulation was used to examine the degree to which speech intelligibility was affected by slow and fast WDRC in conjunction with the presence or absence of DNR. The WDRC and DNR settings were chosen to be reasonable estimates of hearing aids currently available to consumers. RESULTS: A WDRC × DNR × Hearing Status interaction was observed, such that DNR was beneficial for speech intelligibility when combined with fast WDRC speeds, but DNR was detrimental to speech intelligibility when WDRC speeds were slow. The pattern of the WDRC × DNR interaction was observed for both listener groups. Significant main effects of reverberation time and SNR were observed, indicating better performance with lower reverberation times and more positive SNR. CONCLUSIONS: DNR reduced low-amplitude noise before WDRC-amplified the low-intensity portions of the signal, negating one potential downside of fast WDRC and leading to an improvement in speech intelligibility in this simulation. These data suggest that, in some real-world environments that include both reverberation and noise, older listeners with hearing impairment may find speech to be more intelligible if DNR is activated when the hearing aid has fast compression time constants. Additional research is needed to determine the appropriate DNR strength and to confirm results in wearable hearing aids and a wider range of listening environments.

Exploration of the In Vitro Violacein Synthetic Pathway with Substrate Analogues.

Evolution has gifted enzymes with the ability to synthesize an abundance of small molecules with incredible control over efficiency and selectivity. Central to an enzyme's role is the ability to selectively catalyze reactions in the milieu of chemicals within a cell. However, for chemists it is often desirable to extend the substrate scope of reactions to produce analogue(s) of a desired product and therefore some degree of enzyme promiscuity is often desired. Herein, we examine this dichotomy in the context of the violacein biosynthetic pathway. Importantly, we chose to interrogate this pathway with tryptophan analogues in vitro, to mitigate possible interference from cellular components and endogenous tryptophan. A total of nine tryptophan analogues were screened for by analyzing the substrate promiscuity of the initial enzyme, VioA, and compared to the substrate tryptophan. These results suggested that for VioA, substitutions at either the 2- or 4-position of tryptophan were not viable. The seven analogues that showed successful substrate conversion by VioA were then applied to the five enzyme cascade (VioABEDC) for the production of violacein, where l-tryptophan and 6-fluoro-l-tryptophan were the only substrates which were successfully converted to the corresponding violacein derivative(s). However, many of the other tryptophan analogues did convert to various substituted intermediaries. Overall, our results show substrate promiscuity with the initial enzyme, VioA, but much less for the full pathway. This work demonstrates the complexity involved when attempting to analyze substrate analogues within multienzymatic cascades, where each enzyme involved within the cascade possesses its own inherent promiscuity, which must be compatible with the remaining enzymes in the cascade for successful formation of a desired product.

Multienzymatic Cascades and Nanomaterial Scaffolding-A Potential Way Forward for the Efficient Biosynthesis of Novel Chemical Products.

Synthetic biology is touted as the next industrial revolution as it promises access to greener biocatalytic syntheses to replace many industrial organic chemistries. Here, it is shown to what synthetic biology can offer in the form of multienzyme cascades for the synthesis of the most basic of new materials-chemicals, including especially designer chemical products and their analogs. Since achieving this is predicated on dramatically expanding the chemical space that enzymes access, such chemistry will probably be undertaken in cell-free or minimalist formats to overcome the inherent toxicity of non-natural substrates to living cells. Laying out relevant aspects that need to be considered in the design of multi-enzymatic cascades for these purposes is begun. Representative multienzymatic cascades are critically reviewed, which have been specifically developed for the synthesis of compounds that have either been made only by traditional organic synthesis along with those cascades utilized for novel compound syntheses. Lastly, an overview of strategies that look toward exploiting bio/nanomaterials for accessing channeling and other nanoscale materials phenomena in vitro to direct novel enzymatic biosynthesis and improve catalytic efficiency is provided. Finally, a perspective on what is needed for this field to develop in the short and long term is presented.

Rapid Assessment of Subjective Hearing Complaints With a Modified Version of the Tinnitus and Hearing Survey.

Hearing difficulties are frequently reported by patients in audiology clinics, including patients with normal audiometric thresholds. However, because all individuals experience some difficulty understanding speech in noisy environments, it can be difficult to assess hearing complaints objectively across patients. Normative values help address this issue by providing an objective cutoff score for determining what is or is not clinically significant. The goal of this study was to establish normative values for the four-item hearing subscale of the Tinnitus and Hearing Survey (THS-H). Respondents completing the THS-H rate the level of difficulty understanding speech in the situations most commonly reported as being difficult: in the presence of noise, on TV or in movies, soft voices and group conversations. In this study, 22,583 US Service Members (SMs) completed the THS-H using an 11-point scale ranging from 0 (not a problem) to 10 (a very big problem). Responses to the four items were summed to produce values between 0 and 40. The distribution of the final scores was analyzed based on severity of hearing loss, age, and sex. Only 5% of SMs with clinically normal hearing scored above 27, so this score was selected as a cutoff for "clinically significant hearing problems." Due to its ease of administration and interpretation, the THS-H could be a useful tool for identifying patients with subjective hearing difficulty warranting audiological evaluation and management.

Bacterial Membrane Vesicles for In Vitro Catalysis.

The use of biological systems in manufacturing and medical applications has seen a dramatic rise in recent years as scientists and engineers have gained a greater understanding of both the strengths and limitations of biological systems. Biomanufacturing, or the use of biology for the production of biomolecules, chemical precursors, and others, is one particular area on the rise as enzymatic systems have been shown to be highly advantageous in limiting the need for harsh chemical processes and the formation of toxic products. Unfortunately, biological production of some products can be limited due to their toxic nature or reduced reaction efficiency due to competing metabolic pathways. In nature, microbes often secrete enzymes directly into the environment or encapsulate them within membrane vesicles to allow catalysis to occur outside the cell for the purpose of environmental conditioning, nutrient acquisition, or community interactions. Of particular interest to biotechnology applications, researchers have shown that membrane vesicle encapsulation often confers improved stability, solvent tolerance, and other benefits that are highly conducive to industrial manufacturing practices. While still an emerging field, this review will provide an introduction to biocatalysis and bacterial membrane vesicles, highlight the use of vesicles in catalytic processes in nature, describe successes of engineering vesicle/enzyme systems for biocatalysis, and end with a perspective on future directions, using selected examples to illustrate these systems' potential as an enabling tool for biotechnology and biomanufacturing.

Enzyme assembly on nanoparticle scaffolds enhances cofactor recycling and improves coupled reaction kinetics.

Enzyme activity can be many times enhanced in configurations where they are displayed on a nanoparticle (NP) and this same format sometimes even provides access to channeling phenomena within multienzyme cascades. Here, we demonstrate that such enhancement phenomena can be expanded to enzymatic cofactor recycling along with the coupled enzymatic processes that they are associated with. We begin by showing that the efficiency of glucose driven reduction of nicotinamide adenine dinucleotide (NAD+ → NADH) by glucose dehydrogenase (GDH) is enhanced ca. 5-fold when the enzyme is displayed on nanocrystalline semiconductor quantum dots (QDs) which are utilized as prototypical NP materials in our experimental assays. Coupling this enzymatic step with NADH-dependent lactate dehydrogenase (LDH) conversion of lactate to pyruvate also increases the latter's rate by a similar amount when both enzymes were jointly incorporated into self-assembled QD-based nanoclusters. Detailed agarose gel mobility assays and transmission electron microscopy imaging studies confirm that both tetrameric enzymes assemble to and crosslink the QDs into structured nanoclusters via their multiple-pendant terminal (His)6 sequences. Unexpectedly, control experiments utilizing blocking peptides to prevent enzyme-crosslinking of QDs resulted in even further enhancement of individual enzyme on-QD kinetic activity. This activity was also probed revealing that 200-fold excess peptide/QD addition enhanced individual GDH and LDH on-QD kcat a further 2- and 1.5×, respectively, above that seen just by QD display to a maximum of ∼10-fold GDH enhancement. The potential implications for how these enzyme kinetics-enhancing phenomena can be applied to single and multi-enzyme cascaded reactions in the context of cofactor recycling and cell-free synthetic biology are discussed.

Different Strategies Affect Enzyme Packaging into Bacterial Outer Membrane Vesicles.

All Gram-negative bacteria are believed to produce outer membrane vesicles (OMVs), proteoliposomes shed from the outermost membrane. We previously separately engineered E. coli to produce and package two organophosphate (OP) hydrolyzing enzymes, phosphotriesterase (PTE) and diisopropylfluorophosphatase (DFPase), into secreted OMVs. From this work, we realized a need to thoroughly compare multiple packaging strategies to elicit design rules for this process, focused on (1) membrane anchors or periplasm-directing proteins (herein "anchors/directors") and (2) the linkers connecting these to the cargo enzyme; both may affect enzyme cargo activity. Herein, we assessed six anchors/directors to load PTE and DFPase into OMVs: four membrane anchors, namely, lipopeptide Lpp', SlyB, SLP, and OmpA, and two periplasm-directing proteins, namely, maltose-binding protein (MBP) and BtuF. To test the effect of linker length and rigidity, four different linkers were compared using the anchor Lpp'. Our results showed that PTE and DFPase were packaged with most anchors/directors to different degrees. For the Lpp' anchor, increased packaging and activity corresponded to increased linker length. Our findings demonstrate that the selection of anchors/directors and linkers can greatly influence the packaging and bioactivity of enzymes loaded into OMVs, and these findings have the potential to be utilized for packaging other enzymes into OMVs.

Self assembling nanoparticle enzyme clusters provide access to substrate channeling in multienzymatic cascades.

Access to efficient enzymatic channeling is desired for improving all manner of designer biocatalysis. We demonstrate that enzymes constituting a multistep cascade can self-assemble with nanoparticle scaffolds into nanoclusters that access substrate channeling and improve catalytic flux by orders of magnitude. Utilizing saccharification and glycolytic enzymes with quantum dots (QDs) as a model system, nanoclustered-cascades incorporating from 4 to 10 enzymatic steps are prototyped. Along with confirming channeling using classical experiments, its efficiency is enhanced several fold more by optimizing enzymatic stoichiometry with numerical simulations, switching from spherical QDs to 2-D planar nanoplatelets, and by ordering the enzyme assembly. Detailed analyses characterize assembly formation and clarify structure-function properties. For extended cascades with unfavorable kinetics, channeled activity is maintained by splitting at a critical step, purifying end-product from the upstream sub-cascade, and feeding it as a concentrated substrate to the downstream sub-cascade. Generalized applicability is verified by extending to assemblies incorporating other hard and soft nanoparticles. Such self-assembled biocatalytic nanoclusters offer many benefits towards enabling minimalist cell-free synthetic biology.

Trophic ontogeny of a generalist predator is conserved across space.

Consumers can influence ecological patterns and processes through their trophic roles and contributions to the flow of energy through ecosystems. However, the diet and associated trophic roles of consumers commonly change during ontogeny. Despite the prevalence of ontogenetic variation in trophic roles of most animals, we lack an understanding of whether they change consistently across local populations and broad geographic gradients. We examined how the diet and trophic position of a generalist marine predator varied with ontogeny across seven broadly separated locations (~ 750 km). We observed a high degree of heterogeneity in prey consumed without evidence of spatial structuring in this variability. However, compound-specific isotope analysis of amino acids revealed remarkably consistent patterns of increasing trophic position through ontogeny across local populations, suggesting that the roles of this generalist predator scaled with its body size across space. Given the high degree of diet heterogeneity we observed, this finding suggests that even though the dietary patterns differed, the underlying food web architecture transcended variation in prey species across locations for this generalist consumer. Our research addresses a gap in empirical field work regarding the interplay between stage-structured populations and food webs, and suggests ontogenetic changes in trophic position can be consistent in generalist consumers.

Alternative design strategies to help build the enzymatic retrosynthesis toolbox.

Most of the complex molecules found in nature still cannot be synthesized by current organic chemistry methods. Given the number of enzymes that exist in nature and the incredible potential of directed evolution, the field of synthetic biology contains perhaps all the necessary building blocks to bring about the realization of applied enzymatic retrosynthesis. Current thinking anticipates that enzymatic retrosynthesis will be implemented using conventional cell-based synthetic biology approaches where requisite native, heterologous, designer, and evolved enzymes making up a given multi-enzyme pathway are hosted by chassis organisms to carry out designer synthesis. In this perspective, we suggest that such an effort should not be limited by solely exploiting living cells and enzyme evolution and describe some useful yet less intensive complementary approaches that may prove especially productive in this grand scheme. By decoupling reactions from the environment of a living cell, a significantly larger portion of potential synthetic chemical space becomes available for exploration; most of this area is currently unavailable to cell-based approaches due to toxicity issues. In contrast, in a cell-free reaction a variety of classical enzymatic approaches can be exploited to improve performance and explore and understand a given enzyme's substrate specificity and catalytic profile towards non-natural substrates. We expect these studies will reveal unique enzymatic capabilities that are not accessible in living cells.

Tunable Electronic Structure via DNA-Templated Heteroaggregates of Two Distinct Cyanine Dyes.

Molecular excitons are useful for applications in light harvesting, organic optoelectronics, and nanoscale computing. Electronic energy transfer (EET) is a process central to the function of devices based on molecular excitons. Achieving EET with a high quantum efficiency is a common obstacle to excitonic devices, often owing to the lack of donor and acceptor molecules that exhibit favorable spectral overlap. EET quantum efficiencies may be substantially improved through the use of heteroaggregates-aggregates of chemically distinct dyes-rather than individual dyes as energy relay units. However, controlling the assembly of heteroaggregates remains a significant challenge. Here, we use DNA Holliday junctions to assemble homo- and heterotetramer aggregates of the prototypical cyanine dyes Cy5 and Cy5.5. In addition to permitting control over the number of dyes within an aggregate, DNA-templated assembly confers control over aggregate composition, i.e., the ratio of constituent Cy5 and Cy5.5 dyes. By varying the ratio of Cy5 and Cy5.5, we show that the most intense absorption feature of the resulting tetramer can be shifted in energy over a range of almost 200 meV (1600 cm-1). All tetramers pack in the form of H-aggregates and exhibit quenched emission and drastically reduced excited-state lifetimes compared to the monomeric dyes. We apply a purely electronic exciton theory model to describe the observed progression of the absorption spectra. This model agrees with both the measured data and a more sophisticated vibronic model of the absorption and circular dichroism spectra, indicating that Cy5 and Cy5.5 heteroaggregates are largely described by molecular exciton theory. Finally, we extend the purely electronic exciton model to describe an idealized J-aggregate based on Förster resonance energy transfer (FRET) and discuss the potential advantages of such a device over traditional FRET relays.

Cobalt(III)- and rhodium(III)-porphyrin complexes for the effective degradation of fentanyl: Biological inactivation and mechanistic insights.

Cobalt(III) and rhodium(III) complexes containing the water-soluble porphyrin ligand meso-tri(4-sulfonatophenyl)mono(4-carboxyphenyl)porphine (C1S3TPP), [Rh(C1S3TPP)]Nax•nH2O (1) and [Co(C1S3TPP)]Nax•nH2O (2) were prepared from the direct reaction of free porphyrin and metal chloride salts in refluxing MeOH/DMF or EtOH/H2O. Compounds 1 and 2 were characterized using UV-vis and 1H NMR spectroscopies, and high-resolution mass spectrometry. Cell culture based assays of opioid receptor activation showed that while the rhodium complex reduced fentanyl opioid activity 113-fold to an IC50 value of 1.7 µM, the cobalt complex reduced fentanyl activity by 160-fold to an IC50 value of 2.4 µM. An oxidative mechanism for fentanyl breakdown is proposed.

Implementing Multi-Enzyme Biocatalytic Systems Using Nanoparticle Scaffolds.

Interest in multi-enzyme synthesis outside of cells (in vitro) is becoming far more prevalent as the field of cell-free synthetic biology grows exponentially. Such synthesis would allow for complex chemical transformations based on the exquisite specificity of enzymes in a "greener" manner as compared to organic chemical transformations. Here, we describe how nanoparticles, and in this specific case-semiconductor quantum dots, can be used to both stabilize enzymes and further allow them to self-assemble into nanocomplexes that facilitate high-efficiency channeling phenomena. Pertinent protocol information is provided on enzyme expression, choice of nanoparticulate material, confirmation of enzyme attachment to nanoparticles, assay format and tracking, data analysis, and optimization of assay formats to draw the best analytical information from the underlying processes.

Updating the Spectral Correlation Index: Integrating Audibility and Band Importance Using Speech Intelligibility Index Weights.

The original Spectral Correlation Index (SCIo ) is a measure of amplitude envelope distortion that has been used in several studies to predict behavioral results. Because the original SCIo did not account for the differential contribution of particular frequency bands to speech intelligibility (i.e., band importance) or for audibility, a new "individual" version (the SCIi ) is proposed and evaluated. Sentence intelligibility data are used to compare the predictive power and goodness-of-fit for statistical models using two versions of the SCI. The SCIi provides significantly better fits to behavioral data than the SCIo . This result demonstrates the importance of accounting for and including signal audibility in analyzing and modeling data collected from the population of individuals with hearing impairment. With this update, the SCIi is a useful measure for predicting speech intelligibility based on amplitude envelope distortions.

Tuning between Quenching and Energy Transfer in DNA-Templated Heterodimer Aggregates.

Molecular excitons, which propagate spatially via electronic energy transfer, are central to numerous applications including light harvesting, organic optoelectronics, and nanoscale computing; they may also benefit applications such as photothermal therapy and photoacoustic imaging through the local generation of heat via rapid excited-state quenching. Here we show how to tune between energy transfer and quenching for heterodimers of the same pair of cyanine dyes by altering their spatial configuration on a DNA template. We assemble "transverse" and "adjacent" heterodimers of Cy5 and Cy5.5 using DNA Holliday junctions. We find that the transverse heterodimers exhibit optical properties consistent with excitonically interacting dyes and fluorescence quenching, while the adjacent heterodimers exhibit optical properties consistent with nonexcitonically interacting dyes and disproportionately large Cy5.5 emission, suggestive of energy transfer between dyes. We use transient absorption spectroscopy to show that quenching in the transverse heterodimer occurs via rapid nonradiative decay to the ground state (∼31 ps) and that in the adjacent heterodimer rapid energy transfer from Cy5 to Cy5.5 (∼420 fs) is followed by Cy5.5 excited-state relaxation (∼700 ps). Accessing such drastically different photophysics, which may be tuned on demand for different target applications, highlights the utility of DNA as a template for dye aggregation.

Self-assembled nanoparticle-enzyme aggregates enhance functional protein production in pure transcription-translation systems.

Cell-free protein synthesis systems (CFPS) utilize cellular transcription and translation (TX-TL) machinery to synthesize proteins in vitro. These systems are useful for multiple applications including production of difficult proteins, as high-throughput tools for genetic circuit screening, and as systems for biosensor development. Though rapidly evolving, CFPS suffer from some disadvantages such as limited reaction rates due to longer diffusion times, significant cost per assay when using commercially sourced materials, and reduced reagent stability over prolonged periods. To address some of these challenges, we conducted a series of proof-of-concept experiments to demonstrate enhancement of CFPS productivity via nanoparticle assembly driven nanoaggregation of its constituent proteins. We combined a commercially available CFPS that utilizes purified polyhistidine-tagged (His-tag) TX-TL machinery with CdSe/CdS/ZnS core/shell/shell quantum dots (QDs) known to readily coordinate His-tagged proteins in an oriented fashion. We show that nanoparticle scaffolding of the CFPS cross-links the QDs into nanoaggregate structures while enhancing the production of functional recombinant super-folder green fluorescent protein and phosphotriesterase, an organophosphate hydrolase; the latter by up to 12-fold. This enhancement, which occurs by an undetermined mechanism, has the potential to improve CFPS in general and specifically CFPS-based biosensors (faster response time) while also enabling rapid detoxification/bioremediation through point-of-concern synthesis of similar catalytic enzymes. We further show that such nanoaggregates improve production in diluted CFPS reactions, which can help to save money and extend the amount of these costly reagents. The results are discussed in the context of what may contribute mechanistically to the enhancement and how this can be applied to other CFPS application scenarios.

Neonatal encephalopathy plasma metabolites are associated with neurodevelopmental outcomes.

BACKGROUND: To investigate mechanisms of injury and recovery in neonatal encephalopathy (NE), we performed targeted metabolomic analysis of plasma using liquid chromatography with tandem mass spectrometry (LC/MS/MS) from healthy term neonates or neonates with NE. METHODS: Plasma samples from the NE (n = 45, day of life 0-1) or healthy neonatal (n = 30, ≥36 weeks gestation) cohorts had LC/MS/MS metabolomic profiling with a 193-plex targeted metabolite assay covering >366 metabolic pathways. Metabolite levels were compared to 2-year neurodevelopmental outcomes measured by the Bayley Scales of Infant and Toddler Development III (Bayley-III). RESULTS: Out of 193 metabolites, 57 met the pre-defined quality control criteria for analysis. Significant (after false discovery rate correction) KEGG (Kyoto Encyclopedia of Genes and Genomes) pathways included aminoacyl-tRNA biosynthesis, arginine biosynthesis, and metabolism of multiple amino acids. Significant disease pathways included seizures. In regression models, histidine and C6 sugar amine were significantly associated with cognitive, motor, and language and betaine with cognitive and motor Bayley-III composite scores. The addition of histidine, C6 sugar amine, and betaine to a Sarnat score-based clinical regression model significantly improved model performance (Akaike information criterion and adjusted r2) for Bayley-III cognitive, motor, and language scores. CONCLUSIONS: Plasma metabolites may help to predict neurological outcomes in neonatal brain injury and enhance current clinical predictors. IMPACT: Plasma metabolites may help to predict neurological outcomes in NE and supplement current clinical predictors. Current metabolomics research is limited in terms of clinical application and association with long-term outcomes. Our study presents novel associations of plasma metabolites from the first 24 h of life and 2-year neurodevelopmental outcomes for infants with NE. Our metabolomics discovery provides insight into possible disease mechanisms and methods to rescue and/or supplement metabolic pathways involved in NE. Our metabolomics discovery of metabolic pathway supplementations and/or rescue mechanisms may serve as adjunctive therapies for NE.

Using Machine Learning and the National Health and Nutrition Examination Survey to Classify Individuals With Hearing Loss.

Even before the COVID-19 pandemic, there was mounting interest in remote testing solutions for audiology. The ultimate goal of such work was to improve access to hearing healthcare for individuals that might be unable or reluctant to seek audiological help in a clinic. In 2015, Diane Van Tasell patented a method for measuring an audiogram when the precise signal level was unknown (patent US 8,968,209 B2). In this method, the slope between pure-tone thresholds measured at 2 and 4 kHz is calculated and combined with questionnaire information in order to reconstruct the most likely audiograms from a database of options. An approach like the Van Tasell method is desirable because it is quick and feasible to do in a patient's home where exact stimulus levels are unknown. The goal of the present study was to use machine learning to assess the effectiveness of such audiogram-estimation methods. The National Health and Nutrition Examination Survey (NHANES), a database of audiologic and demographic information, was used to train and test several machine learning algorithms. Overall, 9,256 cases were analyzed. Audiometric data were classified using the Wisconsin Age-Related Hearing Impairment Classification Scale (WARHICS), a method that places hearing loss into one of eight categories. Of the algorithms tested, a random forest machine learning algorithm provided the best fit with only a few variables: the slope between 2 and 4 kHz; gender; age; military experience; and self-reported hearing ability. Using this method, 54.79% of the individuals were correctly classified, 34.40% were predicted to have a milder loss than measured, and 10.82% were predicted to have a more severe loss than measured. Although accuracy was low, it is unlikely audibility would be severely affected if classifications were used to apply gains. Based on audibility calculations, underamplification still provided sufficient gain to achieve ~95% correct (Speech Intelligibility Index ≥ 0.45) for sentence materials for 88% of individuals. Fewer than 1% of individuals were overamplified by 10 dB for any audiometric frequency. Given these results, this method presents a promising direction toward remote assessment; however, further refinement is needed before use in clinical fittings.